Harnessing engineered T regulatory cells to promote beta cell health in T1D

利用工程化 T 调节细胞促进 1 型糖尿病 (T1D) 中 β 细胞的健康

• 批准号: 10605317
• 负责人: Jane Hoyt Buckner
• 金额: $ 97.57万
• 依托单位: BENAROYA RESEARCH INST AT VIRGINIA MASON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-08 至 2026-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10605317
• 关键词: Address; Adult; Affect; Antigen Presentation; Antigens; Autoimmune Diseases; Autoimmune Responses; Autoimmunity; Beta Cell; CD4 Positive T Lymphocytes; CD8B1 gene; Cell Survival; Cell physiology; Cells; Cellular Stress; Child; Clinical; Cytoprotection; Disease; Elements; Engineered Gene; Engineering; Environment; FOXP3 gene; Functional disorder; Genes; Goals; Health; Home; Homing; Human; Hyperglycemia; Immune Targeting; Immunosuppression; Inflammation; Inflammatory; Insulin; Insulin-Dependent Diabetes Mellitus; Intervention; Islets of Langerhans; Knowledge; Measures; Mediating; Organ; Pancreas; Prevention; Recovery; Regulatory T-Lymphocyte; Research Personnel; Risk; Signal Transduction; Site; Specificity; Stress; Suppressor-Effector T-Lymphocytes; T-Cell Activation; T-Cell Receptor; T-Lymphocyte; Testing; Therapeutic; Time; Work; antigen test; autoimmune pathogenesis; biological adaptation to stress; cell growth; cell injury; constitutive expression; deamidation; design; draining lymph node; effector T cell; endoplasmic reticulum stress; engineered T cells; immunoengineering; islet; islet cell antibody; knowledge integration; lentivirally transduced; mouse model; neoantigens; novel; peripheral blood; repaired; targeted delivery; targeted treatment; therapeutic protein; tissue injury; tissue repair

Summary Type 1 diabetes (T1D) is characterized by the destruction of β-cells, driven through autoimmune attack directed at the pancreatic islet. In concert with β-cell destruction, β-cell stress and injury contribute to disease by initiating enzymatic activities that compromise β-cell function and generate neoantigens. The investigators of this project will integrate knowledge of clinical T1D, β-cell stress, islet reactive T cells, regulatory T cells (Tregs) and gene engineering to create Tregs that will home to the islet and suppress autoimmunity and inflammation in a manner that will create an environment that will allow β-cell recovery and promote β-cell health. These engineered Tregs (EngTregs) will be generated utilizing homology-directed repair (HDR)-based gene editing to mediate constitutive expression of FOXP3 combined with lentiviral transduction of T cell receptor (TCR) sequences specific to antigens presented in the islet during periods of β-cell stress. Further, we propose develop EngTregs with the ability to co-deliver a payload (with or without TCR engagement) that will function to suppress ongoing inflammation and/or promote β-cell survival and growth. Aim 1 will generate stressed-islet-specific CD4+ and CD8+ EngTregs via gene editing and, in parallel, confer specificity to the pancreatic islet via expression of TCRs that recognize citrullinated or deamidated β-cell-derived neoepitopes. The central hypothesis for Aim 1 is that targeting neoantigens generated during periods of β-cell stress will enhance the targeted delivery of EngTregs to the site of tissue injury. Neoepitope discovery studies will identify TCR sequences restricted to novel β-cell stress neoepitopes. Functional studies will assess stressed-islet-specific EngTregs activation in the presence of β-cell stress as well as in the presence of CD4+ and CD8+ effector T cells. Aim 2 will generate islet-specific CD4+ and CD8+ EngTregs that mediate targeted immune suppression and co-deliver an islet-protective therapeutic cargo. In parallel, we will develop EngTregs that release their cargo upon TCR recognition of antigen. The proposed studies integrate primary human cell studies with relevant T1D murine models, facilitating more rapid identification of promising candidate engineered T cell products. The proposed studies are directly responsive to RFA-DK-21-005 with its request to 1) engineer antigen-specific Treg cells that can home to the pancreatic islet or pancreatic draining lymph nodes and inhibit effector T cells in these compartments; 2) engineer islet- homing synthetic suppressor cells, such as CD4+ T cells engineered to locally produce factors that dampen inflammation, inactivate effector T cells, or promote islet tissue repair; and 3) engineer T cells that produce factors with trophic effects on β-cells to promote function, immunoprotection and/or replication.

概括 1型糖尿病（T1D）的特征是通过针对自身免疫攻击的β细胞破坏 在胰岛。与β细胞破坏一致，β细胞应力和损伤通过引发疾病有助于疾病 损害β细胞功能并产生新抗原的酶促活性。该项目的调查人员 将整合临床T1D，β细胞应力，胰岛反应性T细胞，调节性T细胞（TREG）和基因的知识 工程制造的Tregs可以回家胰岛并以某种方式抑制自身免疫性和注射 这将创造一个环境，该环境将允许β细胞恢复并促进β细胞健康。这些设计的Tregs （Engtregs）将使用基于同源的维修（HDR）基因编辑来生成以调解一致性 FOXP3的表达与T细胞受体（TCR）序列的慢病毒转移相结合 在β细胞应激期间，在胰岛中呈现的抗原。此外，我们建议与 能够连接有效载荷（有或没有TCR参与）的能力，以抑制正在进行的 炎症和/或促进β细胞存活和生长。 AIM 1将产生应力特异性CD4+和 CD8+ Engtregs通过基因编辑，并同时通过TCR表达对胰岛的会议特异性 这可以识别柠檬奶粉或脱膜的β细胞衍生的新垂体。目标1的中心假设是 靶向在β细胞应力期间产生的新抗原将增强靶向递送 到组织损伤部位。 NeoEpitope Discovery研究将识别仅限于新型β细胞的TCR序列 压力卵子。功能研究将在存在的情况下评估应力特异性的Engtregs激活 β细胞应力以及CD4+和CD8+效应T细胞的存在。 AIM 2将生成胰岛特异性CD4+ 和CD8+ Engtregs介导靶向免疫抑制并共同替代胰岛保护疗法 货物。同时，我们将开发出在TCR识别抗原的情况下释放其货物的Engtregs。 拟议的研究将原代人细胞研究与相关的T1D鼠模型整合在一起，支持更快 确定应许的候选工程T细胞产品。拟议的研究直接响应 向RFA-DK-21-005提出要求1）可以回家胰腺的工程师抗原特异性Treg单元 胰岛或胰腺排干淋巴结和抑制这些隔室的效应T细胞； 2）工程师iSlet- HOTRING合成抑制细胞，例如为局部产生该死的因素而设计的CD4+ T细胞 炎症，灭活效应T细胞或促进胰岛组织修复； 3）生产的工程师T细胞 对β细胞产生营养作用以促进功能，免疫保护和/或复制的因素。